Acquisition of a Confocal Laser Scanning Microscope for Cell Biology and Biophysics

采购用于细胞生物学和生物物理学的共聚焦激光扫描显微镜项目摘要伍兹霍尔海洋生物实验室(MBL)的Peter J.S.Smith博士已获得赠款，用于购买具有多光子激发能力的共聚焦激光扫描显微镜和图像分析系统。该系统将被安置在MBL中央显微镜设施中，向包括伍兹霍尔海洋研究所的科学家、访问和暑期调查人员以及在MBL提供的许多研究生和博士后教育课程在内的广泛的常驻研究人员社区提供该系统。调查小组已经制定了从基础生物学到生物医学的几个拟议的研究项目。在MBL的居民社区中，这些包括：1.对锥虫体内囊泡贩运的检查。作为处理传染病的更大项目的一部分。钙/钙调素激酶II在长时程增强中的作用研究记忆基础的更大项目的一部分；3.单细胞真核极端微生物的生理学。研究适应性进化和蛋白质表达/修饰的更大项目的一部分。病原菌的多重耐药性。5.有丝分裂纺锤体的构筑动力学。5.细胞动态实时成像的持续项目。神经元中的钙振荡。这是几个正在进行的项目的一个组成部分，该项目着眼于细胞膜和细胞溶质中的钙活动区域。环境污染物对神经元发育的影响。毒理学领域中MBL调查人员正在研究的一个关键问题。红系细胞中铁运输的机制，这构成了在MBL启动的新的线粒体计划的一部分。MBL的科学家长期以来一直对细胞动力学的研究感兴趣，这必然需要对生命系统进行观察。所选的显微镜最大限度地发挥了这种能力。选择系统组件是为了提供1)可见光光谱中不同荧光团的共聚焦荧光成像的能力；2)多光子激发；3)发射指纹技术，用于同时记录具有近距离发射光谱的多个荧光团；4)足够多的物镜，以便能够对活细胞的结构研究和实验；5)荧光标记结构的精确定位；6)对研究中的活细胞进行温度控制；7)软件，以充分利用共焦的数据采集能力；以及8)图像分析和显示软件。拨款中建议的研究活动反映了MBL社区对科学兴趣的多样性，以及在研究和教育方面的推广。新的显微镜将是用于尖端研究的MBL设施的无价补充。它对居民和游客的可用性(每年有200多名季节性调查人员出于研究目的访问MBL)将确保授予的赠款具有很高的影响力和用途。

Acquisition of a Confocal Laser Scanning Microscope for Cell Biology and BiophysicsProject SummaryA grant has been awarded to Dr Peter J.S. Smith of the Marine Biological Laboratory (MBL), Woods Hole, for the acquisition of a confocal laser scanning microscope and image analysis system with multiphoton excitation capability. The system is to be housed in the MBL Central Microscopy Facility, making it available to a wide community of resident researchers, including scientists from the Woods Hole Oceanographic Institute, visiting and summer investigators, as well as by many of the graduate- and postdoctoral-level education courses offered at the MBLSeveral proposed research projects have already been formulated by the investigating team, spanning basic biology to the biomedical sciences. Amongst the resident community of the MBL these include:1. An examination of vesicle trafficking in trypanosomes. Part of a larger project dealing infectious diseases.2. The role of Ca/CaM kinase II dynamics in LTP. Part of a larger project investigating the foundations of memory; 3. The physiology of single-celled eukaryotic extremophiles. Part of a larger project to investigate adaptive evolution and protein expression/modification.4. Multidrug resistance in pathogens. 5. Architectural dynamics of the mitotic spindle. A continuing project on live imaging of cellular dynamics.6. Calcium oscillations in neurons. A component of several ongoing projects looking at calcium activity domains in the cell membrane and cytosol.7. The effects of environmental pollutants on neuronal development. A key issue being studied by MBL investigators in the field of toxicology.8. The mechanisms of iron trafficking in erythroid cells which forms part of a new mitochondrial program initiated at the MBL. Scientists at the MBL have had a long interest in the study of cellular dynamics, which by necessity requires observations on living systems. The microscope to be chosen maximizes this ability. System components were chosen to provide 1) capability for confocal fluorescent imaging of different fluorophores in the visible spectrum; 2) multiphoton excitation; 3) emission fingerprinting for simultaneous recording of multiple fluorophores with closely-spaced emission spectra; 4) a sufficient range of objective lenses to enable both structural studies and experiments on the physiology of living cells; 5) precisely localization fluorescent-labeled structures; 6) temperature control of live cells under study; 7) software to use fully the data acquisition capabilities of the confocal; and 8) software for image analysis and display. The research activities proposed in the grant reflect the diversity of scientific interest amongst the MBL community as well as outreach in research and education. The new microscope will be an invaluable addition the MBL facilities for cutting edge research. Its availability to residents and visitors alike (over 200 seasonal investigators visit the MBL for research purposes on an annual basis) will insure a high impact and use for the grant awarded.